S-1-propenyl-L-cysteine suppresses lipopolysaccharide-induced expression of matrix metalloproteinase-1 through inhibition of tumor necrosis factor-α converting enzyme-epidermal growth factor receptor axis in human gingival fibroblasts

Hiroshi Nango; Masahiro Ohtani

doi:10.1371/journal.pone.0284713

Peer Review History

Original SubmissionOctober 6, 2022
9 Feb 2023 Decision Letter - Bruno Giros, Editor PONE-D-22-27612S-1-propenyl-L-cysteine suppresses lipopolysaccharide-induced expression of matrix metalloproteinase-1 through inhibition of tumor necrosis factor-α converting enzyme-epidermal growth factor receptor axis in human gingival fibroblastsPLOS ONE Dear Dr. Nango, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. Please submit your revised manuscript by Mar 26 2023 11:59PM. If you will need more time than this to complete your revisions, please reply to this message or contact the journal office at plosone@plos.org. 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We note that you have stated that you will provide repository information for your data at acceptance. Should your manuscript be accepted for publication, we will hold it until you provide the relevant accession numbers or DOIs necessary to access your data. If you wish to make changes to your Data Availability statement, please describe these changes in your cover letter and we will update your Data Availability statement to reflect the information you provide. [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Partly ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: The introduction part presents a good introduction to the topic and the literature is relevant. The research is methodologically correctly appointed, and the presentation of the results (tables, figures) corresponds to the objectives of the research and the set hypothesis. The discussion is extensive and debates the set goal and results of the study. Altogether, the paper is well designed and well written. Reviewer #2: The manuscript by Hango. Et al. describes the impact of S1PC (a bioactive compound derived from aged garlic extract) on HGF1 cells. The authors observed decreased mRNA expression, intracellular levels, and secretion of MMP1 after the treatment. They linked this reduction with the inhibition of LPS-induced phosphorylation of EGFR and cleavage of TACE. Overall, the manuscript depicts a straightforward hypothesis based on the group's extensive experience with aged garlic extracts using a simple experimental design. Major issues: Mostly related to the explanation of sample size and technical replicas. In all figures, the number of replicas is relatively low (n=3-5), and it is unclear if that number depicts separate experiments or just replicas within a single experiment. Further, the low number of replicas impacted the statistical differences between groups with clear tendencies that are not significant (i.e., Figure 2a at 72h, figure 2b at 30uM between no LPS and LPS, and so on). Given the experimental design’s simplicity (not primary cell culture, but a cell line), the number of replicas should be high enough to guarantee a precise statistical analysis. Also, I suggest using all bar graphs the individual values to understand high SD. Minor issues: Since 1998 (ref 2), a significant understanding of the pathogenesis of periodontal disease has been achieved in a way that is no longer considered a “chronic infection,” as stated in the abstract, or caused by pathogenic bacteria (line 29). Instead, Periodontitis is a chronic multifactorial inflammatory disease associated with dysbiotic plaque biofilms (https://doi.org/10.1002/JPER.17-0721). Figure 1 b, has a duplicated group (LPS+, S1PC+). ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No ******** [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. 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Revision 1
16 Feb 2023 Author Response Point-by-point responses to the Editor and Reviewers (Manuscript ID: PONE-D-22-27612) We are grateful to the Editor and Reviewers for evaluating our manuscript and for providing us with such valuable comments. Below, please find our point-by-point responses to each comment; the comments made by the reviewer are in black font, and our responses are in blue font. The revised areas in the manuscript are marked with red text. Reviewer #1 The introduction part presents a good introduction to the topic and the literature is relevant. The research is methodologically correctly appointed, and the presentation of the results (tables, figures) corresponds to the objectives of the research and the set hypothesis. The discussion is extensive and debates the set goal and results of the study. Altogether, the paper is well designed and well written. Response: We appreciate your kind comment. Reviewer #2: The manuscript by Hango. Et al. describes the impact of S1PC (a bioactive compound derived from aged garlic extract) on HGF1 cells. The authors observed decreased mRNA expression, intracellular levels, and secretion of MMP1 after the treatment. They linked this reduction with the inhibition of LPS-induced phosphorylation of EGFR and cleavage of TACE. Overall, the manuscript depicts a straightforward hypothesis based on the group's extensive experience with aged garlic extracts using a simple experimental design. Response: We appreciate your kind comment and hope that we were able to adequately address all your concerns. Your comments and suggestions have greatly improved our manuscript. Major issues: Mostly related to the explanation of sample size and technical replicas. In all figures, the number of replicas is relatively low (n=3-5), and it is unclear if that number depicts separate experiments or just replicas within a single experiment. Response: We have missed to clarify details for the replica. The number of replicates in each figure caption represents the number of independent experiments. We have added details about the number of replicates to each figure caption in the revised manuscript. In addition, we are sorry to have made careless mistakes for the number of experiments. The incorrect sample sizes in Fig. 1, 3a, 4a, and Fig. S1 in the original manuscript have been corrected in the revised manuscript. Further, the low number of replicas impacted the statistical differences between groups with clear tendencies that are not significant (i.e., Figure 2a at 72h, figure 2b at 30uM between no LPS and LPS, and so on). Given the experimental design’s simplicity (not primary cell culture, but a cell line), the number of replicas should be high enough to guarantee a precise statistical analysis. Response: We agree that this point is important from a statistical viewpoint. As the reviewer pointed out, it has been reported that the statistical power increases with an increase in sample size (Naegle K et al., Sci. Signal., 2015), so we expect statistically significant differences in our clear trend data with increasing sample size. However, we consider, at least, that our data sufficiently support the beneficial effects of S1PC with statistical significance (LPS alone vs. LPS + S1PC) even at the current sample size. We also think that our sample size (n = 3－6) is not particularly low in general, considering that several articles using cell lines recently published in PLOS ONE were conducted with two to four independent experiments (Maniscalco E et al., Plos One, 2023; Sharif M et al., Plos One, 2023; Shi Y et al., Plos One, 2023). This suggestion is controversial as a general issue of determining an appropriate sample size in in vitro experiments and should be given proper attention in future experiments. Also, I suggest using all bar graphs the individual values to understand high SD. Response: Overall, we agree with you on this point. We added individual values as dot plots to each graph in the revised manuscript. Minor issues: Since 1998 (ref 2), a significant understanding of the pathogenesis of periodontal disease has been achieved in a way that is no longer considered a “chronic infection,” as stated in the abstract, or caused by pathogenic bacteria (line 29). Instead, Periodontitis is a chronic multifactorial inflammatory disease associated with dysbiotic plaque biofilms (https://doi.org/10.1002/JPER.17-0721). Response: Thank you for your important suggestion. We have modified these sentences (line 2－3 in the abstract section and line 27－29 in the introduction section) and replaced reference 2 to the following suggested reference in the revised manuscript. 2. Papapanou PN, Sanz M, Buduneli N, Dietrich T, Feres M, Fine DH, et al. Periodontitis: Consensus report of workgroup 2 of the 2017 World Workshop on the Classification of Periodontal and Peri-Implant Diseases and Conditions. J Periodontol. 2018;89: S173–S182. doi:10.1002/JPER.17-0721 Figure 1 b, has a duplicated group (LPS+, S1PC+). Response: Done as suggested. Attachments Attachment Submitted filename: Responcse to Reviewers.docx https://doi.org/10.1371/journal.pone.0284713.r002
6 Apr 2023 Decision Letter - Bruno Giros, Editor S-1-propenyl-L-cysteine suppresses lipopolysaccharide-induced expression of matrix metalloproteinase-1 through inhibition of tumor necrosis factor-α converting enzyme-epidermal growth factor receptor axis in human gingival fibroblasts PONE-D-22-27612R1 Dear Dr. Nango, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Bruno Giros, Ph.D. Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #2: All comments have been addressed ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #2: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #2: All comments have been satisfactory addressed. The author corrected the minor issues and specified the number of independent experiments performed. ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review? For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #2: No ******** https://doi.org/10.1371/journal.pone.0284713.r003
Formally Accepted
13 Apr 2023 Acceptance Letter - Bruno Giros, Editor PONE-D-22-27612R1 S-1-propenyl-L-cysteine suppresses lipopolysaccharide-induced expression of matrix metalloproteinase-1 through inhibition of tumor necrosis factor-α converting enzyme-epidermal growth factor receptor axis in human gingival fibroblasts Dear Dr. Nango: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Professor Bruno Giros Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0284713.r004

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